For optimal ChIP and ChIP-seq results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.
Application
Dilution
Western Blotting
1:1000
Immunoprecipitation
1:50
Immunofluorescence (Immunocytochemistry)
1:400 - 1:1600
Flow Cytometry (Fixed/Permeabilized)
1:50 - 1:200
Chromatin IP
1:50
Storage
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.
For a carrier free (BSA and azide free) version of this product see product #76431.
Specificity / Sensitivity
Tri-Methyl-Histone H3 (Lys9) (D4W1U) Rabbit Monoclonal Antibody detects endogenous levels of histone H3 when tri-methylated on Lys9. This antibody shows some cross-reactivity with histone H3 that is di-methylated on Lys9, but does not cross-react with non-methylated or mono-methylated histone H3 Lys9. This antibody does not detect tri-methyl histone H3 Lys9 when the adjacent Ser10 residue is phosphorylated during mitosis. In addition, this antibody does not cross-react with methylated histone H3 Lys4, Lys27, Lys36, or Lys79.
Species Reactivity:
Human, Mouse, Rat, Monkey
Species predicted to react based on 100% sequence homology:
Bovine
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of histone H3 in which Lys9 is tri-methylated.
